TWI422584B - Composition for treating cancer cells and use therefor - Google Patents

Description

Composition for killing cancer cells and use thereof

The present invention relates to a composition for killing cancer cells and a preparation method thereof, and more particularly to a composition containing an extract component of a plant of the genus Tubocapsicum spp. and a preparation method thereof.

Although today's medical technology is developing rapidly, there are still no fundamental treatments for diseases such as cancer, cardiovascular disease and even AIDS. Because the therapeutic drugs used in Western medicine often lead to many side effects, alternative therapies including Chinese herbal medicine have become the subject of popular research. Many plant-derived extracts and their derivatives, such as vinblastine (vincristinem and vinblastine), camptothecin (camptothecin), and paclitaxel (taxol) have been widely used in the treatment of tumors.

Among them, Dragon Ball " Tubocapsicum anomalum (Franch. & Sav.) Makino" belongs to the Solanaceae Tubocapsicum plant, which is mainly distributed in Southeast Asia and is about 0-2,000 meters above sea level. In Taiwan, this plant is considered as folk medicine for the treatment of gonorrhea, dysentery, nephritis and swollen poison. As for its effectiveness against cell poisoning, it is still not clear.

In view of this, the inventors have conceived the case in the light of the long-term careful experiment and research, and the spirit of perseverance, in view of the above-mentioned shortcomings of the prior art. The following is a brief description of the case.

The purpose of this case is to provide an extract of the Solanaceae plant Dragon Ball ( Tubocapsicum anomalum ) containing unique compounds withanolides for human hepatoma cells (Hep G2, Hep 3B), lung cancer cells (A549) and breast cancer cells. (MDA-MB-231, MCF-7) has a cytotoxic effect.

The present invention provides a composition for killing cancer cells, which comprises a compound of the form of the formula (withanolides) of the formula:

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells comprising a compound withanolides having the structure shown in Formula 2:

Wherein R ₁ is selected from the group consisting of a hydroxyl group and a C1-C4 alkoxy group.

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells, which comprises a compound of the form of the asanolides having the structure shown in Formula 3:

Wherein R ₂ is selected from one of hydrogen, a hydroxyl group, a halogen and a C1-C4 alkoxy group.

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells comprising a compound withanolides having the structure shown in Formula 4:

Wherein R ₃ is a hydroxyl group.

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells comprising a compound withanolides having the structure shown in Formula 5:

Wherein R ₅ is selected from one of a hydroxyl group and a C1-C4 alkoxy group.

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells comprising a compound of the form of the formula (withanolides) of the formula:

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells, which comprises a compound of the form of the asanolides having the structure shown in Formula 7:

Wherein R ₆ is selected from one of hydrogen and hydroxyl.

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells comprising a compound withanolides having the structure shown in Formula 8:

Wherein R ₇ and R ₉ are each selected from one of hydrogen and a hydroxyl group, and R ₈ is selected from one of hydrogen, a hydroxyl group and a halogen.

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells comprising a compound withanolides having the structure shown in Formula 10:

Wherein R ₁₀ and R ₁₁ are each selected from one of hydrogen and hydroxyl.

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells comprising a compound withanolides having the structure shown in Formula 11:

Wherein R ₁₂ and R ₁₃ are each selected from one of hydrogen and hydroxyl.

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a composition for killing cancer cells comprising a compound withanolides having the structure shown in Formula 12:

Preferably, the withanolides compound is obtained by extracting from a Solanaceae plant.

Preferably, the Solanaceae plant is Tubocapsicum anomalum .

The present invention provides a method for preparing the compound of the formula (withanolides) according to Formulas I to 12, comprising the steps of: providing a Dragon Ball ( Tubocapsicum anomalum ), and extracting the Dragon Ball with a first organic solution, A first extract is obtained. The first extract is then extracted with a second organic solution to obtain a second extract. The second extract is then extracted with a third organic solution to obtain a third extract. Finally, the second and third extracts are separated to obtain the withanolides-like compound.

Preferably, the first organic solution is methanol.

Preferably, the second organic solution is ethyl acetate.

Preferably, the third organic solution is n-butanol.

Preferably, the second and the third extraction are separated by chromatography. Take the material to obtain the compound withanolides.

According to the composition provided above and the preparation method thereof, the present invention not only obtains a new lycopene compound from a natural plant, but more particularly, the novel lycopene compound has a bacterium that kills cancer cells. active.

The other objects, features, and advantages of the invention will be apparent from the description and appended claims. These examples are merely illustrative and should not be construed as limiting the scope of the invention.

The present invention discloses a composition for killing cancer cells and a preparation method thereof, the contents of which will be described below by way of preferred embodiments, but the embodiments are only preferred, and the practice of the present invention is not limited to such In the preferred embodiment, other embodiments may be devised by those skilled in the art, and the embodiments are intended to be within the scope of the invention.

The composition provided by the present invention comprises a compound withanolides, which is an extract obtained from a plant of Tubocapsicum anomalum , which is then provided to prepare the extract of the present invention. A preferred embodiment of the method.

Embodiment 1

The collected Dragon Ball ( Tubocapsicum anomalum ) whole grass 2.5 kg, with methanol 3 liters repeatedly extracted five times to obtain a first extract, concentrated under reduced pressure to dry brown viscous material, with ethyl acetate and water oscillated Separation. After the ethyl acetate layer was concentrated under reduced pressure, a second extract was obtained. Next, the n-butanol was separated from the water by shaking, and the n-butanol layer was concentrated under reduced pressure to obtain a third extract. The ethyl acetate layer was column of chromatography (Si gel, 230 ~ 400mesh, 5 20cm ×) further separated to _{n -Hexane → n -Hexane / CHCl 3} → CHCl 3 → CHCl 3 / MeOH → MeOH In order to extract the solvent system, it is judged to be roughly divided into 16 fractions by the thin layer chromatography analysis (TLC).

Part 8 (Fr. 8, 529.8 mg) was eluted from the ethyl acetate layer with CHCl ₃ : MeOH = 20:1. The crystals were found to be crystallised, and the crystals were washed by MeOH to give the dansyl lactone A (Tubocapsanolide A, 42.5). Mg, CHCl ₃ : MeOH = 20:1, R _f = 0.5).

Part 11 (Fr. 11, 165.2 mg) was eluted from the ethyl acetate layer with CHCl ₃ : MeOH = 15:1. The crystals were found to be crystallized, and the crystals were washed by filtration with MeOH to give the tribendone lactone A (Tubocapsenolide A). 101.1 mg, CHCl ₃ : MeOH = 25:1, R _f = 0.2).

The 12th part (Fr. 12, 120.1 mg) was extracted from the ethyl acetate layer with CHCl ₃ : MeOH = 15:1. The crystals were found to be crystallized, and the crystals were washed with MeOH to give ananonolide C (Anomanolide C, 65.3 mg, CHCl ₃ : MeOH = 10:1, R _f = 0.5).

Part 8 to Part 12 were combined to obtain sample one (TAEw, 400 mg), and the mixture was extracted in a ratio of H ₂ O: MeOH: CHCl ₃ = 1: 4: 5 to obtain a methanol layer (250 mg) and a chloroform layer (70 mg). . The methanol layer (TAEwm) was subjected to high performance liquid chromatography (HPLC) to [ODS 250 × 21.2 mm, MeOH: H ₂ O = 65: 35, PDA-detector (UV-230 nm), flow rate: The ratio of 3 mL/min] was purified and separated into 6 fractions. Part 1-4 is Tubocapsenolide A [41 mg, ODS 250 x 21.2 mm, MeOH: H2O = 65:35, flow rate: 3 mL/min, Rt . = 28 min] , 1-6 Pearl steroidal enol lactone moiety is A (Tubocapsanolide A, 10mg, ODS 250 × 21.2mm, MeOH: H 2 O = 65: 35, flow rate: 3mL / min, Rt = 35min ).

Part 1-2 (Fr.1-2,106.86mg) using high performance liquid chromatography (HPLC) to [ODS 250 × 10mm, MeOH: H 2 O = 50: 50, PDA-detector (UV-230nm), The ratio of flow rate: 2 mL/min] was purified and separated into 6 fractions.

Section 1-2-3 (Fr. 1-2-3, 8.63 mg) using high performance liquid chromatography (HPLC) to [ODS 250 x 10 mm, MeOH: H ₂ O = 45: 55, PDA-detector Purification and separation (UV-230 nm), flow rate: 2 mL/min], wherein the fraction 1-2-3-2 was obtained as Anomanolide E [3.5 mg, ODS 250 × 4.6, MeOH: H ₂ O = 45: 55, flow rate: 1 mL/min, Rt = 11.4 min.

Part 1-2-4 (23 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 10 mm, MeOH: H ₂ O = 45: 55, PDA-detector (UV-230 nm), flow rate (flow) Purification and separation in a ratio of 2 mL/min], wherein the 1-2-4-3 part is ananonolide A (Anomanolide A, 14 mg, ODS 250×4.6 mm, MeOH: H ₂ O=45) : 55, flow rate: 1 mL/min, Rt = 12.4 min).

Part 1-3 (32 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 10 mm, MeOH: H ₂ O = 45: 55, PDA-detector (UV-230 nm), flow rate: 2 mL/min Purification and separation were carried out in which the fractions 1-3-4 were obtained as Tubocapsenolide B [2.3 mg, ODS 250×10 mm, MeOH: H ₂ O=45:55, flow rate: 2 mL] /min, Rt = 62.5min].

After the 1-5 (26 mg) was dissolved in methanol, it was allowed to stand for several days, and a white solid was precipitated. Filtration with cotton was carried out to obtain Tubocapsenolide D [20 mg, ODS 250 × 4.6 mm, MeOH: H ₂ O = 50:50, flow rate: 1 mL/min, Rt = 24 min].

The n-butanol layer (5.9 g) was subjected to extraction with a ratio of H ₂ O: CHCl ₃ = 1:1 to obtain a chloroform layer (TABC 1.38 g) and an aqueous layer (TABH 4.2 g). The chloroform layer (TABC) was determined by column chromatography (Sephadex LH-20, 4.5×50 cm), MeOH was used as the solvent solvent system, and judged by thin layer chromatography (TLC). (fractions).

Part 4 (276.5mg) was subjected to column chromatography (Si gel, 230~400mesh, 2.5×27cm) with full CHCl ₃ as the solvent solvent system, sequentially increasing the polarity to MeOH and subdividing into 21 parts ( Fractions).

Section 4-5 (17.15 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 10 mm, MeOH: H ₂ O = 75: 25, UV-detector (UV-230 nm), flow rate (flow rate) ): Purification and separation in a ratio of 2 mL/min], wherein Part 4-05 is obtained as Tubocapsanolide C [2.7 mg, ODS 250×10 mm, MeOH: H ₂ O=75:25, flow rate ( Flow rate): 2 mL/min, Rt = 42.5 min].

Section 4-10 (6.22 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 4.6 mm, MeOH: H ₂ O = 70:30, PDA-detector (UV-230 nm), flow rate (flow) Purification and separation of the ratio: 1 mL/min], wherein the parts 4-10 were obtained from Tubocapsenolide E [2.7 mg, ODS 250×4.6 mm, MeOH: H ₂ O=70: 30, flow rate: 1 mL/min, Rt = 14.3 min].

Part 4-17 (11.1 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 4.6 mm, MeOH: H ₂ O = 70:30, PDA-detector (UV-230 nm), flow rate (flow) Rate): Purification and separation of the ratio of 1 mL/min], wherein the 4-17-1 part was obtained from Tubocapsenolide F [5.12 mg, ODS 250×4.6 mm, MeOH: H ₂ O= 70:30, flow rate: 1 mL/min, Rt = 17.5 min].

Part 5 (278.3mg) was subjected to column chromatography (Si gel, 230~400mesh, 2×25cm) with CHCl ₃ : MeOH=150:1 as the solvent solvent system, sequentially increasing the polarity to MeOH, subdividing In 22 fractions, the fraction 5-02 was obtained as a tuna caprolactone E (Tubocapsanolide E, 15.6 mg, CHCl ₃ : MeOH = 20:1, R _f = 0.5).

Section 5-03 (2.47 mg) was performed using a high performance liquid chromatography (HPLC) [ODS 250 x 10 mm, MeOH: H ₂ O = 70:30, PDA-detector (UV-230 nm), flow rate: 1 mL/ The ratio of min] was purified and separated, wherein the fraction 5-03 was borneol lactone D (Tubocapsanolide D, 1.2 mg, ODS 250×10 mm, MeOH: H ₂ O=70:30, flow rate: 2 mL/min, Rt =17.4min).

Part 5-07 (39.38 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 4.6 mm, MeOH: H ₂ O = 70:30, PDA-detector (UV-230 nm), flow rate: 1 mL The ratio of /min] was purified and isolated, wherein part 5-07 was obtained as ananonolide B (Anomanolide B, 8.3 mg, ODS 250×4.6 mm, MeOH: H ₂ O=70:30, flow rate: 1 mL/ Min, Rt = 6.1 min).

Section 5-09 (15.7 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 10 mm, MeOH: H ₂ O = 60: 40, PDA-detector (UV-230 nm), flow rate: 2 mL / Purification and separation of the ratio of min], wherein the 5-9 part was obtained as a pipercapto lactone B (Tubocapsanolide B, 8.3 mg, ODS 250×10 mm, MeOH: H ₂ O=60:40, flow rate: 2 mL/min , Rt = 40.8 min).

Section 5-11 (19.66mg) using high performance liquid chromatography (HPLC) to [ODS 250 × 10mm, MeOH: H 2 O = 65: 35, PDA-detector (UV-230nm), flow rate: 2mL / The ratio of min] was purified and purified, wherein fr. 5-11 was obtained as Tubocapsenolide G [11.53 mg, ODS 250×4.6 mm, MeOH: H ₂ O=65:35, flow rate (flow) Rate): 1 mL/min, Rt = 4.1 min].

Part 5-18 (15.15 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 10 mm, MeOH: H ₂ O = 65: 35, PDA-detector (UV-230 nm), flow rate (flow rate) ): Purification and separation was carried out at a ratio of 2 mL/min], wherein the fraction 5-18-1 was obtained as Tubocapsenolide C [6.72 mg, ODS 250×10 mm, MeOH: H ₂ O=65: 35, flow rate: 2 mL / min, Rt = 17.6 min].

Part 6 (108.73mg) was subjected to column chromatography (Si gel, 230~400mesh, 2×22cm) with CHCl ₃ : MeOH=50:1 as the solvent solvent system, sequentially increasing the polarity to MeOH, subdividing In 16 fractions, parts 6-12 were obtained from Anomanolide D [33.5 mg, CHCl ₃ : MeOH = 10:1, R _f = 0.4].

Sections 6-14 (2.05 mg) using high performance liquid chromatography (HPLC) with [ODS 250 x 4.6 mm, MeOH: H ₂ O = 50: 50, PDA-detector (UV-230 nm), flow rate (flow) Rate): 1 mL/min] was purified and separated, wherein the fractions 6-14-4 were obtained as Tubanolide A [1.15 mg, ODS 250×4.6 mm, MeOH: H ₂ O=50:50 Flow rate: 1 mL/min, Rt = 13.8 min].

Embodiment 2

The collected fresh Dragon Ball root [ Tubocapsicum anomalum (Franch. & Sav.) Makino] was continuously extracted five times with methanol for 24 hours. The extract was concentrated under reduced pressure and then extracted with water and ethyl acetate. The ethyl acetate layer and the aqueous layer were collected, and the aqueous layer was subjected to partition extraction with n-butanol to obtain an aqueous layer and an n-butanol layer; and the ethyl acetate layer was subjected to extraction with methanol and n-hexane to collect a methanol layer and a n-hexane layer. . The methanol layer was separated by column chromatography (Sephadex LH-20, 1.6×28 cm), and the MeOH was used as a solvent solvent system, subdivided into 5 fractions, and the third fraction (807 mg) was used as a column. The chromatographic method (Si gel, 230~400 mesh, 5×20 cm) was further separated, and CHCl ₃ →CHCl ₃ /MeOH→MeOH was used as the solvent solvent system, and judged by thin layer chromatography (TLC). Subdivided into 28 fractions.

Part 3-3 uses high performance liquid chromatography (HPLC) to [ODS 250 x 10 mm, MeOH: H ₂ O = 65: 35, UV/VIS-detector (UV-230 nm), flow rate: The ratio of 2 mL/min] was purified and separated into 8 fractions. The first part is obtained wherein 3-3-7 iso sterol lactone Pearl G (iso -tubocapsanolide G) [ODS 250 × 10mm, MeOH: H 2 O = 65: 35, UV / VIS-detector (UV-230nm), flow rate (flow rate): 2mL/min, Rt=34min]

Part 3-3-3 uses high performance liquid chromatography (HPLC) to [ODS 250×10 mm, MeOH: H ₂ O=65:35, UV/VIS-detector (UV-230 nm), flow rate (flow rate) ): 2 mL/min] recycling to obtain the 3-3-3-1 part as 20-hydroxy-tubocapsanolide A [ODS 250×10 mm, MeOH: H ₂ O=65:35, UV/VIS-detector (UV-230nm), flow rate: 2mL/min, Rt=17min] and part 3-3-3-2 is 20-hydroxy-dragonol Ester G (20-hydroxy-tubocapsanolide G) [ODS 250×10 mm, MeOH: H ₂ O=65:35, UV/VIS-detector (UV-230 nm), flow rate: 2 mL/min, Rt=18 min ].

Section 3-3-4 uses high performance liquid chromatography (HPLC) to [ODS 250 x 10 mm, MeOH: H ₂ O = 65: 35, UV/VIS-detector (UV-230 nm), flow rate (flow rate) ): 2 mL / min] recycling to obtain the 3-3-4-1-2 part of the tuna caprolactone H (Tubocapsanolide H) [ODS 250 × 10mm, MeOH: H ₂ O = 65: 35 , UV/VIS-detector (UV-230nm), flow rate: 2mL/min, Rt=24min], Section 3-3-4-2 is Tubocapsanolide F [ODS 250× 10mm, MeOH: H ₂ O=65:35, UV/VIS-detector (UV-230nm), flow rate: 2mL/min, Rt=25min] and Section 3-3-4-4 is Dragon Ball sterol Lactone G (Tubocapsanolide G) [ODS 250 x 10 mm, MeOH: H ₂ O = 65:35, UV/VIS-detector (UV-230 nm), flow rate: 2 mL/min, Rt = 26 min].

Sections 3-8 utilize high performance liquid chromatography (HPLC) to [ODS 250 x 10 mm, MeOH: H ₂ O = 65: 35, UV/VIS-detector (UV-230 nm), flow rate: Purification and separation were carried out at a ratio of 2 mL/min], wherein the part 3-8-2 was obtained as 23-Hydroxy-tubocapsanolide A [ODS 250×10 mm, MeOH: H ₂ O=65 : 35, UV/VIS-detector (UV-230 nm), flow rate: 2 mL/min, Rt = 13.5 min].

The first 3-10 (126.8mg) in a column of chromatography (Si gel, 230 ~ 400mesh, 3 × 15cm), in CHCl _3: MeOH = 20: 1 The elution solvent system is subdivided into three parts (fractions), taking part 3-10-3 (11mg) using high performance liquid chromatography (HPLC) to [ODS 250×10mm, MeOH: H ₂ O=60:40, UV/VIS-detector (UV) -230 nm), flow rate: 2 mL/min] was purified and separated, wherein the 3-10-3-2 fraction was obtained as Anomanolide F [ODS 250×10 mm, MeOH: H ₂ O = 60: 40, UV/VIS-detector (UV-230 nm), flow rate: 2 mL/min, Rt = 14.5 min].

The preferred embodiment of the method of the present invention for preparing the extract is described above. According to the extraction method of this case, the properties of the twenty-five new pure compounds with tonallides obtained from Tubocapsicum anomalum are further identified as follows:

About the physicochemical properties of the compound Tubocapsenolide A(1):

Melting point (mp): 223-225 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 212, 228 nm; infrared absorption spectrum (IR): ν _max 3380, 2921, 1677, 1380, 1130 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, pyridine- d ₅ ): δ 6.47 (1H, d, J = 10.0 Hz, H-2 ), 7.27 (1H, dd, J = 10.0, 6.4 Hz, H-3 ) , 4.05 (1H, d, J = 6.4 Hz, H-4 ), 3.37 (1H, s(br), H-6 ), 1.39 (1H, m, H-7a ), 2.33 (1H, m, H- 7b ), 2.35 (1H, m, H-8 ), 1.37 (1H, m, H-9 ), 1.26 (1H, ddd, J = 12.4, 4.8, 4.0 Hz, H-11a ), 2.04 (1H, m , H-11b ), 1.77 (1H, m, H-12a ), 2.04 (1H, m, H-12b ), 2.48 (2H, m, H-15a, b ), 4.45 (1H, t, J = 8.0 Hz, H-16 ), 1.21 (3H, s, CH ₃ -18 ), 1.81 (3H, s, CH ₃ -19 ), 2.58 (1H, qd, J = 7.2, 3.6 Hz, H-20 ), 1.17 (3H,d, J = 7.2 Hz, CH ₃ -21 ), 5.17 (1H, ddd, J = 12.8, 3.6, 3.2 Hz, H-22 ), 2.47 (1H, m, H-23a ), 2.30 (1H , m, H-23b ), 1.87 (3H, s, CH ₃ -27 ), 1.77 (3H, s, CH ₃ -28 ).

Mass Spectrum EI-MS m / z ( s. s.): 469 [M+H] ⁺ .

High-resolution mass spectrometry HR-ESI-MS: m / z 469.2594 [M+H] ⁺ (calc. C ₂₈ H ₃₇ O ₆ , 469.2585).

About the physicochemical properties of the compound Tubocapsenolide B(2):

Melting point (mp): 133-135 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 208, 228 nm; infrared absorption spectrum (IR): ν _max 3421, 2921, 1691, 1380, 1130 cm ^-1 Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, pyridine- d ₅ ): δ 2.50 (1H, m, H-2a ), 2.79 (1H, J = 15.2, 8.0, 7.6 Hz, H-2b ), 1.96 (1H) , m, H-3a ), 2.22 (1H, m, H-3b ), 3.54 (1H, dd, J = 4.0, 3.6 Hz, H-4 ), 3.24 (1H, s(br), H-6 ) , 1.25 (1H, m, H-7a ), 2.35 (1H, ddd, J = 14.4, 4.0, 2.0 Hz, H-7b ), 2.10 (1H, m, H-8 ), 1.40 (1H, dd, J =12.4, 11.2 Hz, H-9 ), 1.06 (1H, m, H-11a ), 2.04 (1H, m, H-11b ), 1.77 (2H, m, H-12a, b ), 2.43 (2H, m, H-15a, b ), 4.12 (1H, t, J = 7.6 Hz, H-16 ), 1.12 (3H, s, CH ₃ -18 ), 1.30 (3H, s, CH ₃ -19 ), 2.16 (1H, qd, J = 7.2, 3.2 Hz, H-20 ), 0.98 (3H, d, J = 7.2 Hz, CH ₃ -21 ), 4.35 (1H, ddd, J = 12.4, 7.2, 3.2 Hz, H -22 ), 2.48 (1H, m, H-23a ), 2.24 (1H, m, H-23b ), 1.87 (3H, s, CH ₃ -27 ), 1.94 (3H, s, CH ₃ -28 ).

Mass spectrum EI-MS m / z ( s. s.): 471 [M+H] ⁺ .

High-resolution mass spectroscopy HR-ESI-MS: m / z 493.2568 [M+Na] ⁺ (calc. C ₂₈ H ₃₈ O ₆ Na, 493.2561).

About the physicochemical properties of the compound Tubocapsenolide C(3):

Melting point (mp): 226-228 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 208, 228 nm; infrared absorption spectrum (IR): ν _max 3411, 2919, 1681, 1135, 1060 cm ^-1 Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, CD ₃ OD ): δ 2.58 (1H, dd, J = 12.4, 2.8 Hz, H-2a ), 3.15 (1H, dd, J = 12.4, 6.0 Hz, H- 2b ), 4.06 (1H, dt, J = 6.0, 2.8 Hz, H- 3 ), 3.31 (1H, d, J = 2.8 Hz, H-4 ), 3.28 (1H, s(br), H-6 ) , 1.41 (1H, d, J = 10.8, 2.4, 1.2 Hz, H-7a ), 2.32 (1H, ddd, J = 10.8, 2.4, 1.2 Hz, H-7b ), 2.07 (1H, br, H-8) ), 1.44 (1H, ddd, J = 9.2, 8.0, 1.2 Hz, H-9 ), 1.05 (1H, dd, J = 10.0, 4.0 Hz, H-11a ), 1.68 (1H, dd, J = 10.0, 1.2 Hz, H-11b ), 1.85 (1H, m, H-12a ), 2.07 (1H, m, H-12b ), 2.20 (1H, m, H-15a ), 2.38 (1H, dd, J = 12.8) , 7.2 Hz, H-15b ), 4.10 (1H, t, J = 6.8 Hz, H-16 ), 1.09 (3H, s, CH ₃ -18 ), 1.20 (3H, s, CH ₃ -19 ), 2.20 (1H, m, H-20 ), 1.02 (3H, d, J = 6.0 Hz, CH ₃ -21 ), 4.70 (1H, ddd, J = 10.4, 3.2, 2.4 Hz, H-22 ), 2.54 (1H ,dd, J = 12.4, 10.4 Hz, H-23a ), 2.24 (1H, dd, J = 12.4, 2.4 Hz, H-23b ), 1.96 (3H, s, CH ₃ -27 ), 1.82 (3H, s , CH ₃ -28 )

Mass spectrum EI-MS m / z ( s. s.): 487 [M+H] ⁺ .

High-resolution mass spectroscopy HR-ESI-MS: m / z: 509.2516 [M+Na] ⁺ (calc. C ₂₈ H ₃₈ O ₇ Na, 509.2510).

About the physicochemical properties of the compound Tubocapsenolide D(4):

Melting point (mp): 124~126 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 210, 228 nm; infrared absorption spectrum (IR): ν _max 3409, 2921, 1691, 1384, 1135, 1097 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, CD ₃ OD ): δ 2.63 (1H, dd, J = 14.8, 4.0 Hz, H-2a ), 3.14 (1H, dd, J = 14.8, 5.2 Hz, H-2b), 3.75 (1H , ddd, J = 4.4,4.0,3.6Hz, H-3), 3.51 (1H, d, J = 3.2Hz, H-4), 3.30 (1H, s (br), H-6 ), 1.25 (1H, m, H-7a ), 2.32 (1H, ddd, J = 14.4, 4.0, 2.4 Hz, H-7b ), 2.07 (1H, m, H-8 ), 1.44 (1H , ddd, J = 10.8, 10.0, 2.0 Hz, H-9 ), 1.05 (1H, dd, J = 12.0, 4.0 Hz, H-11a ), 2.02 (1H, m, H-11b ), 1.83 (2H, m, H-12a, b ), 2.42 (2H, m, H-15a, b ), 4.13 (1H, dd, J = 8.4, 7.6 Hz, H-16 ), 1.12 (3H, s, CH ₃ -18 ), 1.30 (3H, s, CH 3 -19), 2.17 (1H, qd, J = 7.2,6.8Hz, H-20), 0.98 (3H, d, J = 7.2Hz, CH 3 -21), 4.37 (1H,ddd, J = 12.8, 6.4, 3.2 Hz, H-22 ), 2.48 (1H, m, H-23a ), 2.30 (1H, dd, J = 17.6, 3.2 Hz, H-23b ), 1.86 ( 3H, s, CH ₃ -27 ), 1.94 (3H, s, CH ₃ -28 ), 3.34 (3H, s, OCH ₃ -1 ' ).

Mass spectrum FAB-MS m / z ( s. s.): 501 [M+H] ⁺ , 523 [M+Na] ⁺ .

High-resolution mass spectroscopy HR-ESI-MS: m / z: 523.2672 [M+Na] ⁺ (calc. C ₂₉ H ₄₀ O ₇ Na, 523.2666).

About the physicochemical properties of the compound Tubocapsenolide E(5):

Melting point (mp): 104-106 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 210, 228 nm; infrared absorption spectrum (IR): ν _max 3413, 2917, 1702, 1687, 1394, 1132 , 1091 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, CDCl ₃ ): δ 2.62 (1H, dd, J = 14.4, 4.0 Hz, H-2a ), 3.14 (1H, dd, J = 14.4, 5.6 Hz , H-2b ), 3.82 (1H, ddd, J = 4.4, 4.4, 4.0 Hz, H-3 ), 3.49 (1H, m, H-4 ), 3.29 (1H, s(br), H-6 ) , 1.23 (1H, J = 14.4, 1.2 Hz, H-7a ), 2.33 (1H, ddd, J = 14.4, 3.2, 2.4 Hz, H-7b ), 2.07 (1H, m(br), H-8 ) , 1.42 (1H, ddd, J = 10.8, 10.0, 1.6 Hz, H-9 ), 1.05 (1H, dd, J = 12.0, 4.4 Hz, H-11a ), 2.02 (1H, m(br), H- 11b ), 1.82 (2H, m, H-12a, b ), 2.42 (2H, m, H-15a, b ), 4.13 (1H, t, J = 8.0 Hz, H-16 ), 1.12 (3H, s _{, CH 3 -18), 1.29 (} 3H, s, CH 3 -19), 2.17 (1H, qd, J = 7.2,6.8Hz, H-20), 0.98 (3H, d, J = 7.2Hz, CH 3 -21 ), 4.37 (1H, ddd, J = 12.8, 6.4, 3.2 Hz, H-22 ), 2.44 (1H, m, H-23a ), 2.20 (1H, dd, J = 17.2, 2.4 Hz, H- 23b ), 1.85 (3H, s, CH ₃ -27 ), 1.93 (3H, s, CH ₃ -28 ), 3.34 (3H, s, OCH ₃ -1 ' ), 3.39 (1H, td, J = 9.2, 6.4 Hz, H-1'a ), 3.50 (1H, td, J = 9.2, 6.4 Hz, H- 1'b ), 1.48 (2H, m, H-2'a, b ), 1.30 (2H, m , H-3'a , b ), 0.88 (3H, t, J = 8.8 Hz, CH ₃ -4' ).

Mass Spectrum ESI-MS m / z ( s. s.): 543 [M+H] ⁺ .

High resolution mass spectrum HR-ESI-MS: m / z 565.3193 [M + Na] + ( calcd for _{C 32 H 46 O 7 Na,} 565.3136).

About the physicochemical properties of the compound Tubocapsenolide F(6):

Melting point (mp): 214~216 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 212, 228 nm; infrared absorption spectrum (IR): ν _max 3424, 2927, 1679, 1380, 1132 cm ^-1 Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.28 (1H, dd, J = 10.0, 2.0 Hz, H-2 ), 6.91 (1H, dd, J = 10.0, 2.0 Hz, H-3 ), 5.37 (1H, s(br), H-4 ), 3.81 (1H, dd, J = 12.8, 4.0 Hz, H-6 ), 2.01 (1H, m, H-7a ), 2.99 ( 1H,dt, J =13.2, 4.0 Hz, H-7b ), 2.30 (1H, m, H-8 ), 1.94 (1H, m, H-9 ), 1.26 (1H, m, H-11a ), 2.01 (1H, m, H-11b ), 1.66 (1H, m, H-12a ), 1.85 (1H, m, H-12b ), 2.38 (2H, m, H-15a,b ), 4.37 (1H, dd , J = 8.0, 7.6 Hz, H-16 ), 1.18 (3H, s, CH ₃ -18 ), 1.73 (3H, s, CH ₃ -19 ), 2.55 (1H, qd, J = 7.6, 3.2 Hz, H-20 ), 1.12 (3H, d, J = 7.6 Hz, CH ₃ -21 ), 5.17 (1H, m, H-22 ), 2.45 (1H, m, H-23a ), 2.30 (1H, dd, J = 14.4, 2.4 Hz, H-23b ), 1.78 (3H, s, CH ₃ -27 ), 1.66 (3H, s, CH ₃ -28 ).

High resolution mass spectrum HR-ESI-MS: m / z 509.2520 [M + Na] + ( calcd for _{C 28 H 38 O 7 Na,} 509.2510).

About the physicochemical properties of the compound Tubocapsenolide G(7):

Melting point (mp): 264~266°C; UV absorption spectrum (UV, in methanol solution): λ _max 216 nm; infrared absorption spectrum (IR): ν _max 3478, 2947, 1723, 1676, 1379, 1126 cm ^-1 Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, CDCl ₃ ): δ 6.01 (1H, dd, J = 10.4, 2.0 Hz, H-2 ), 6.47 (1H, dd, J = 10.4, 2.0 Hz, H-3 ), 5.04 (1H, t, J = 2.0 Hz, H-4 ), 4.45 (1H, dd, J = 10.4, 3.6 Hz, H-6 ), 1.85 (1H, J = 10.4, 10.4 Hz, H-7a) ), 2.51 (1H, ddd, J = 10.4, 3.6, 3.6 Hz, H-7b ), 2.33 (1H, m, H-8 ), 1.50 (1H, td, J = 9.6, 1.6 Hz, H-9 ) , 1.28 (1H, m, H-11a ), 2.01 (1H, m, H-11b ), 1.63 (1H, m, H-12a ), 1.85 (1H, m, H-12b ), 2.40 (1H, m , H-15a ), 2.55 (1H, m, H-15b ), 4.09 (1H, dd, J = 7.6, 6.0 Hz, H-16 ), 1.06 (3H, s, CH ₃ -18 ), 1.20 (3H _{, s, CH 3 -19),} 2.15 (1H, qd, J = 7.2,7.2Hz, H-20), 0.96 (3H, d, J = 7.2Hz, CH 3 -21), 4.37 (1H, ddd, J = 12.8, 7.2, 2.4 Hz, H-22 ), 2.42 (1H, m, H-23a ), 2.22 (1H, dd, J = 18.0, 2.4 Hz, H-23b ), 1.86 (3H, s, CH ₃ -27 ), 1.92 (3H, s, CH ₃ -28 ).

Mass Spectrum ESI-MS m / z ( s. s.): 505 [M+H] ⁺ .

High resolution mass spectrum HR-ESI-MS: m / z 527.2177 [M + Na] + ( calcd for _{C 28 H 37 ClO 6 Na,} 527.2171).

About the physicochemical properties of the compound Tubocapsanolide A (8):

Melting point (mp): 233 ~ 235 ℃ ; ultraviolet absorption spectrum (UV, in methanol): _max 218nm λ; infrared absorption spectrum (IR): ν _max 3403,2918,1688,1679,1380,1132cm ^-1 Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.43 (1H, d, J = 9.6 Hz, H-2 ), 7.23 (1H, dd, J = 9.6, 6.0 Hz, H- 3 ), 4.01 (1H, dd, J = 6.0, 4.6 Hz, H-4 ), 3.20 (1H, s(br), H-6 ), 1.17 (1H, ddd, J = 14.8, 11.2, 1.2 Hz, H-7a ), 2.01 (1H, m, H-7b ), 1.63 (1H, ddd, J = 11.2, 11.2, 4.0 Hz, H-8 ), 0.98 (1H, ddd, J = 11.6, 11.2, 4.0 Hz) , H-9 ), 1.57 (1H, m, H-11a ), 2.01 (1H, m, H-11b ), 1.40 (1H, m, H-12a ), 1.60 (1H, m, H-12b ), 1.24 (1H, ddd, J = 12.0, 11.6, 6.4 Hz, H-14 ), 1.12 (1H, dd, J = 12.4, 12.0 Hz, H-15a ), 1.86 (1H, dd, J = 12.4, 6.4 Hz) , H-15b ), 3.59 (1H, s(br), H-16 ), 0.89 (3H, s, CH ₃ -18 ), 1.81 (3H, s, CH ₃ -19 ), 2.45 (1H, qd, J = 8.8, 6.8 Hz, H-20 ), 1.02 (3H, d, J = 6.8 Hz, CH ₃ -21 ), 3.91 (1H, ddd, J = 12.8, 8.8, 3.2 Hz, H-22 ), 2.22 (1H, dd, J = 17.6, 12.4 Hz, H-23a ), 2.08 (1H, dd, J = 17.6, 3.2 Hz, H-23b ), 1.92 (3H, s, CH ₃ -27 ), 1.73 (3H) , s, CH ₃ -28 ).

Mass spectrum FAB-MS m / z (rel.int.): 469 [M+Na] ⁺ .

High resolution mass spectrum HR-ESI-MS: m / z 469.2594 [M + Na] + ( calcd for _{C 28 H 36 O 6 Na,} 469.2585).

About the physicochemical properties of compound 20-Hydroxytubocapsanolide A(9):

Melting point (mp): 245~247 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 214 nm; infrared absorption spectrum (IR): ν _max 3439, 2922, 2856, 1705, 1380, 1236, 1026 , 750 cm ^-1 ; hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.46 (1H, d, J = 9.5 Hz, H-2 ), 7.22 (1H, dd, J = 6.0 Hz, H-3 ), 4.00 (1H, d, J = 6.0 Hz, H-4 ), 3.21 (1H, br s, H-6 ), 1.16 (1H, m, H-7a ), 2.00 (1H, qd, J =15.0, 2.0 Hz, H-7b ), 1.66 (1H, m, H-8 ), 0.87 (1H, m, H-9 ), 1.60 (1H, m, H-11a ), 1.96 (1H, m , H-11b ), 1.85 (1H, m, H-12a ), 1.93 (1H, m, H-12b ), 1.37 (1H, td, J = 6Hz, H-14 ), 1.10(1H,td, J =13.0, 12.0 Hz, H-15a ), 1.77 (1H, m, H-15b ), 3.57 (1H, s, H-16 ), 1.06 (3H, s, H-18 ), 1.81 (3H, s, H-19 ), 1.45 (3H, s, H-21 ), 4.48 (1H, dd, J = 12.5, 3.5 Hz, H-22 ), 2.17 (1H, dd, J = 18.0, 3.5 H-23a ), 2.83 (1H, t, J = 18.0 Hz, H-23b ), 1.91 (3H, s, H-27 ), 1.74 (3H, s, H-28 ).

Mass Spectrum ESI-MS m / z ( s. s.): 507 [M+Na] ⁺ , 507 (100).

High-resolution mass spectroscopy HR-ESI-MS: 507.2361 [M+Na] ⁺ (calc. C ₂₈ H ₃₆ O ₇ Na, 507.2359).

About the physicochemical properties of compound 23-Hydroxytubocapsanolide A (10):

Melting point (mp): 223~225 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 216 nm; infrared absorption spectrum (IR): ν _max 3409, 2922, 2848, 1690, 1447, 1380, 753 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.42 (1H, d, J = 9.6 Hz, H-2 ), 7.21. (1H, dd, J = 9.6, 6.4 Hz, H-3 ), 3.99 (1H, d, J = 6.4 Hz, H-4 ), 3.18 (1H, br s, H-6 ), 1.16 (1H, m, H-7a ), 1.98 (1H, m, H-7b ), 1.63(1H,m, H-8 ), 0.92(1H,m, H-9 ), 2.01(2H,m, H-11ab ), 1.38(1H,m, H-12a ),1.56 (1H, m, H-12b ), 1.22 (1H, m, H-14 ), 1.13 (1H, m, H-15a ), 1.72 (1H, dd, J = 12.4, 5.6 Hz H-15b ), 3.813 (1H, s, H-16 ), 0.82 (3H, s, H-18 ), 1.79 (3H, s, H-19 ), 2.59 (1H, qd, J = 7.2, 6.8 Hz, H-20 ), 1.15 (3H, d, J = 6.8 Hz, H-21 ), 4.36 (1H, m, H-22 ), 4.39 (1H, d, H-23 ), 1.98 (3H, s, H-27 ), 2.09 (3H, d, J = 0.8Hz , H-28).

Mass Spectrum ESI-MS m / z ( rel. s.): 507 [M+Na] ⁺ , 507 (100), 413 (75), 381 (48), 353 (12).

High-resolution mass spectroscopy HR-ESI-MS: 507.2361 [M+Na] ⁺ (calc. C ₂₈ H ₃₆ O ₇ Na, 507.2359).

About the physicochemical properties of the compound Tubocapsanolide B (11):

Melting point (mp): 225-227 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 207, 223 nm; infrared absorption spectrum (IR): ν _max 3411, 2908, 1698, 1382, 1126 cm ^-1 ; proton nuclear magnetic resonance spectroscopy ^{_{1 H NMR (400MHz, C 5}} D 5 N): δ 2.97 (1H, dd, J = 15.2,3.2Hz, H-2a), 3.15 (1H, dd, J = 15.2,8.0Hz, H -2b ), 3.93 (1H, m, H-3 ), 3.89 (1H, s(br), H-4 ), 3.36 (1H, s(br), H-6 ), 1.32 (1H, m, H -7a ), 2.11 (1H, m, H-7b ), 1.61 (1H, m, H-8 ), 1.32 (1H, m, H-9 ), 1.53 (1H, m, H-11a,b ), 1.39 (1H, m, H-12a ), 1.59 (1H, m, H-12b ), 1.28 (1H, m, H-14 ), 1.14 (1H, dd, J = 12.8, 12.0 Hz, H-15a ) , 1.80 (1H, dd, J = 12.8, 6.0 Hz, H-15b ), 3.60 (1H, s(br), H-16 ), 0.87 (3H, s, CH ₃ -18 ), 1.69 (3H, s , CH ₃ -19 ), 2.45 (1H, qd, J = 8.0, 6.8 Hz, H-20 ), 1.02 (3H, d, J = 6.8 Hz, CH ₃ -21 ), 3.93 (1H, m, H- 22 ), 2.22 (1H, dd, J = 17.2, 12.0 Hz, H-23a ), 2.09 (1H, dd, J = 17.2, 3.2 Hz, H-23b ), 1.86 (3H, s, CH ₃ -27 ) , 1.92 (3H, s, CH ₃ -28 ), 3.33 (3H, s, OCH ₃ -1 ' ).

High resolution mass spectrum HR-ESI-MS: m / z 523.2672 [M + Na] + ( calcd for _{C 29 H 40 O 7 Na,} 523.2676).

About the physicochemical properties of the compound Tubocapsanolide C (12):

Melting point (mp): 223~225 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 208,226 nm; infrared absorption spectrum (IR): ν _max 3388, 2931, 1702, 1687, 1380, 1095 cm ^{- 1} ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, CDCl ₃ ): δ 2.60 (1H, dd, J = 15.2, 3.2 Hz, H-2a ), 2.94 (1H, dd, J = 15.2, 6.4 Hz, H- 2b ), 3.77 (1H, ddd, J = 6.4, 3.2, 2.8 Hz, H- 3 ), 3.49 (1H, d, J = 2.8 Hz, H-4 ), 3.21 (1H, s(br), H- 6 ), 1.35 (1H, m, H-7a ), 2.18 (1H, m, H-7b ), 1.44 (1H, m, H-8 ), 1.19 (1H, td, J = 12.4, 8.0 Hz, H -9 ), 1.46 (1H, m, H-11a, b ), 1.43 (1H, m, H-12a ), 1.60 (1H, m, H-12b ), 1.16 (1H, m, H-14 ), 1.26(1H,d, J =12.0Hz, H-15a ), 1.86(1H,m, H-15b ), 3.46(1H,S(br), H-16 ),0.84(3H,s, CH ₃ - 18 ), 1.29 (3H, s, CH ₃ -19 ), 2.42 (1H, qd, J = 8.8, 7.2 Hz, H-20 ), 0.99 (3H, d, J = 7.2 Hz, CH ₃ -21 ), 3.86 (1H, ddd, J = 12.0, 8.8, 3.6 Hz, H-22 ), 2.30 (1H, dd, J = 19.2, 12.0 Hz, H-23a ), 2.15 (1H, m, H-23b ), 1.86 (3H, s, CH ₃ -27 ), 1.92 (3H, s, CH ₃ -28 ), 3.39 (1H, dt, J = 8.8, 6.4 Hz, H- 1 'a ), 3.48 (1H, dt, J =8.8, 6.4 Hz, H-1'b ), 1.48 (2H, m, H-2' ), 1.34 (2H, m, H-3' ), 0.91 (3H, t, J = 7.2 Hz, CH ₃ -4' ).

Mass Spectrum ESI-MS m / z ( s. s.): 543 [M+H] ⁺ .

High-resolution mass spectroscopy HR-ESI-MS: m / z: 565.3137 [M+Na] ⁺ (calc. C ₃₂ H ₄₆ O ₇ Na, 565.3136).

About the physicochemical properties of the compound Tubocapsanolide D (13):

Melting point (mp): 178-180 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 220 nm; infrared absorption spectrum (IR): ν _max 3491, 2937, 1687, 1382, 1132 cm ^-1 ; nuclear magnetic resonance spectroscopy ^{_{1 H NMR (400MHz, CDCl 3}} ): δ 6.03 (1H, dd, J = 10.0,2.4Hz, H-2), 6.53 (1H, dd, J = 10.0,2.4Hz, H-3), 5.02 (1H, s(br), H-4 ), 4.43 (1H, dd, J = 12.8, 4.8 Hz, H-6 ), 1.64 (1H, m, H-7a ), 2.01 (1H, ddd, J = 9.6, 8.8, 4.8 Hz, H-7b ), 1.63 (1H, d, J = 8.8 Hz, H-8 ), 1.32 (1H, m, H-9 ), 1.01 (1H, m, H-11a ) , 1.56(1H,m, H-11b ), 1.37(1H,m, H-12a ),1.56(1H,m, H-12b ), 1.26(1H,m, H-14 ), 1.36(1H,m , H-15a ), 1.58 (1H, m, H-15b ), 1.43 (1H, m, H-16a ), 2.27 (1H, m, H-16b ), 0.85 (3H, s, CH ₃ -18 ) , 1.26 (3H, s, CH ₃ -19 ), 2.21 (1H, qd, J = 7.2, 1.6 Hz, H-20 ), 1.08 (3H, d, J = 7.2 Hz, CH ₃ -21 ), 4.61 ( 1H,ddd, J = 12.8, 3.2, 1.6 Hz, H-22 ), 2.50 (1H, dd, J = 18.0, 12.8 Hz, H-23a ), 2.36 (1H, dd, J = 18.0, 3.2 Hz, H -23b ), 1.85 (3H, s, CH ₃ -27 ), 1.91 (3H, s, CH ₃ -28 ).

Mass spectrum FAB-MS m / z ( rel. int.): 489 [M+H] ⁺ .

High resolution mass spectrum HR-ESI-MS: m / z 511.2668 [M + Na] + ( calcd for _{C 28 H 40 O 7 Na,} 511.2666).

About the physicochemical properties of the compound Tubocapsanolide F(14):

Melting point (mp): 200~202 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 214 nm; infrared absorption spectrum (IR): ν _max 3453, 2922, 1682, 1376, 1129, 750 cm ^-1 Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.36 (1H, d, J = 10.0 Hz, H-2 ), 7.17 (1H, dd, J = 9.6, 6.4 Hz, H- 3 ), 3.99 (1H, d, J = 6.0, 3.2 Hz, H-4 ), 3.20 (1H, br s, H-6 ), 1.23 (1H, m, H-7a ), 2.11 (1H, m, H-7b ), 1.60 (1H, m, H-8 ), 1.01 (1H, m, H-9 ), 1.71 (1H, m, H-11a ), 2.10 (1H, m, H-11b ), 1.08 (1H, m, H-12a ), 1.66 (1H, m, H-12b ), 1.86 (1H, m, H-14 ), 1.75 (1H, m, H-15a ), 1.95 (1H, m, H -15b ), 1.89 (1H, m, H-16a ), 1.98 (1H, m, H-16b ), 0.73 (3H, s, H-18 ), 1.88 (3H, s, H-19 ), 2.31 ( 1H, qd, J =13.6, 6.8, 2.6 Hz, H-20 ), 1.19 (3H, d, J = 6.8 Hz, H-21 ), 4.76 (1H, td, J = 12.8, 9.6, 3.2 Hz, H -22 ), 2.42 (1H, t, J = 18.0, 12.8 H-23a ), 2.63 (1H, dd, J = 18.0, 3.2, 2.8 Hz, H-23b ), 1.93 (3H, s, H-27 ) , 1.66 (3H, s, H-28 ).

Mass Spectrum ESI-MS m / z (rel. int.): 493 [M+Na] ⁺ , 493 (100), 453 (12), 413 (30), 381 (65), 353 (16).

High resolution mass spectrum HR-ESI-MS: m / z 493.2564 [M + Na] + ( calcd for _{C 28 H 38 O 6 Na,} 493.2566).

About the physicochemical properties of the compound Tubocapsanolide G (15):

Melting point (mp): 218-220 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 206, 230 nm; infrared absorption spectrum (IR): ν _max 3439, 2922, ¹⁶⁹⁰ , 1096, 750 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 2.87 (1H, dd, J = 15.5, 3.5 Hz, H-2a ), 3.08 (1H, dd, J = 15.5, 7.5 Hz, H -2b ), 3.92 (1H, m, H-3 ), 3.90 (1H, br s, H-4 ), 3.37 (1H, br s, H-6 ), 1.37 (1H, m, H-7a ), 2.21 (1H, dd, J = 4.5,2.5Hz, H-7b), 1.61 (1H, m, H-8), 1.34 (1H, m, H-9), 1.08 (1H, m, H-11a) , 1.72(1H,m, H-11b ), 1.73(1H,m, H-12a ), 1.97(1H,m, H-12b ), 1.92(1H,m, H-14 ),1.57(1H,m , H-15a ), 1.61 (1H, m, H-15b ), 1.89 (1H, m, H-16a ), 1.98 (1H, m, H-16b ), 0.73 (3H, s, CH ₃ -18 ) , 1.76 (3H, s, CH ₃ -19 ), 2.34 (1H, dq, J = 7.0, 3.0 Hz, H-20 ), 1.20 (3H, d, J = 7.0 Hz, CH ₃ -21 ), 4.77 ( 1H, td, J =13.0, 3.5, 3.0 Hz, H-22 ), 2.44 (1H, t, J = 18.0, 13.0 Hz, H-23a ), 2.65 (1H, dd, J = 18.0, 3.5 Hz, H -23b ), 1.94 (3H, s, CH ₃ -27 ), 1.65 (3H, s, CH ₃ -28 ), 3.29 (3H, s, OCH ₃ -1 ' ).

Mass Spectrum ESI-MS m / z ( s. s.): 525 [M+Na] ⁺ , 525 (100), 413 (29).

High-resolution mass spectroscopy HR-ESI-MS: m / z 525.2830 [M+Na] ⁺ (calc. C ₂₉ H ₄₂ O ₇ Na, 525.2828).

About the physicochemical properties of the compound iso -Tubocapsanolide G (16):

Melting point (mp): 233-235 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 207, 223 nm; infrared absorption spectrum (IR): ν _max 3424, 2929, 1705, 1384, 1096, 753 cm ^{- 1} ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 2.97 (1H, dd, J = 15.2, 3.2 Hz, H-2a ), 3.15 (1H, dd, J = 15.2, 8.0 Hz , H-2b ), 3.95 (1H, m, H-3 ), 3.92 (1H, d, J = 2.4Hz, H-4 ), 3.41 (1H, s(br), H-6 ), 1.40 (1H , m, H-7a ), 2.19 (1H, m, H-7b ), 1.56 (1H, m, H-8 ), 1.28 (1H, m, H-9 ), 1.50 (1H, m, H-11a ), 1.54 (1H, m, H-11b ), 1.23 (1H, m, H-12a ), 1.96 (1H, m, H-12b ), 1.85 (1H, m, H-14 ), 1.11 (1H, m, H-15a), 1.61 (1H, m, H-15b), 1.75 (1H, m, H-16a), 2.18 (1H, m, H-16b), 0.82 (1H, m, H-17) , 0.97 (3H, s, CH ₃ -18 ), 1.71 (3H, s, CH ₃ -19 ), 1.39 (3H, s, CH ₃ -21 ), 4.38 (1H, dd, J = 12.8, 3.6 Hz, H-22 ), 2.27 (1H, m, H-23a ), 2.54 (1H, m, H-23b ), 1.93 (3H, s, CH ₃ -27 ), 1.81 (3H, s, CH ₃ -28 ) , 3.37 (3H, s, OCH ₃ -1' ).

Mass Spectrum ESI-MS m / z ( s. s.): 525 [M+Na] ⁺ , 525 (100), 413 (12).

High-resolution mass spectroscopy HR-ESI-MS: m / z 525.2830 [M+Na] ⁺ (calc. C ₂₉ H ₄₂ O ₇ Na, 525.2828).

For the physicochemical properties of the compound 20-Hydroxytubocapsanolide G (17):

Melting point (mp): 208-210 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 206, 228 nm; infrared absorption spectrum (IR): ν _max 3461, 2922, 1705, 1384, 1089, 753 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 2.91 (1H, dd, J = 15.2, 3.2 Hz, H-2a ), 3.09 (1H, dd, J = 15.2, 8.0 Hz, H-2b ), 3.91 (1H, m, H-3 ), 3.90 (1H, d, J = 2.4 Hz, H-4 ), 3.37 (1H, br s, H-6 ), 1.39 (1H, m, H-7a ), 2.19 (1H, m, H-7b ), 1.65 (1H, m, H-8 ), 1.44 (1H, m, H-9 ), 1.64 (1H, m, H-11a ) , 1.69 (1H, m, H-11b ), 1.90 (1H, m, H-12a ), 2.16 (1H, m, H-12b ), 2.03 (1H, dq, J = 7.6, 3.6 Hz, H-14 ), 1.17 (1H, m, H-15a ), 1.70 (1H, m, H-15b ), 2.16 (1H, m, H-16a ), 2.99 (1H, m, J = 12.0, 3.2, 2.8 Hz, H-16b ), 1.20 (3H, s, CH ₃ -18 ), 1.76 (3H, s, CH ₃ -19 ), 1.53 (3H, s, CH ₃ -21 ), 5.01 (1H, m, H-22 ), 2.43 (1H, dd, J = 18.0, 3.2 Hz, H-23a ), 2.73 (1H, m, J = 18.0, 14.8 Hz, H-23b ), 1.85 (3H, s, CH ₃ -27 ), 1.67 (3H, s, CH ₃ -28 ), 3.30 (3H, s, OCH ₃ -1 ' ).

Mass Spectrum ESI-MS m / z ( s. s.): 541 [M+Na] ⁺ , 541 (100), 413 (53), 381 (55), 353 (19).

High resolution mass spectrum HR-ESI-MS: m / z 541.2780 [M + Na] + ( calcd for _{C 29 H 42 O 7 Na,} 541.2777).

About the physicochemical properties of the compound Tubocapsanolide H (18):

Melting point (mp): 242 ~ 244 ℃ ; ultraviolet absorption spectrum (UV, in methanol): _max 205,226nm λ; infrared absorption spectra _{(IR): ν max 3431,2922,2856,1705,} 1376,1093, 750 cm ^-1 ; hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 2.97 (1H, dd, J = 15.6, 4.0 Hz, H-2a ), 3.11 (1H, dd, J = 15.6, 8.0 Hz, H-2b ), 3.93 (1H, dd, J = 8.0, 4.0, 3.2 Hz, H- 3 ), 3.90 (1H, d, J = 3.2 Hz, H-4 ), 3.40 (1H, br s , H-6 ), 1.39 (1H, m, H-7a ), 2.18 (1H, m, H-7b ), 1.42 (1H, m, H-9 ), 1.75 (1H, m, H-11a ), 2.18 (1H, m, H-11b ), 1.63 (1H, m , H-12a ), 2.20 (1H, m, H-12b ), 1.50 (1H, dq, H-14 ), 1.85 (1H, m, H-15a), 2.05 (1H , m, H-15b), 6.04 (1H, d, J = 2.0Hz, H-16), 1.03 (3H, s, CH 3 -18), 1.73 (3H, s, CH ₃ -19 ), 1.55 (3H, s, CH ₃ -21 ), 4.55 (1H, dd, J = 12.8, 3.6 Hz, H-22 ), 2.34 (1H, dd, J = 18.0, 3.6 Hz, H -23a ), 2.82 (1H, t, J = 18.0Hz, H-23b ), 1.91 (3H, s, CH ₃ -27 ), 1.74 (3H, s, CH ₃ -28 ), 3.35 (3H, s, OCH ₃ -1' ).

Mass Spectrum ESI-MS m / z ( s. s.): 523 [M+Na] ⁺ , 523 (100), 425 (11), 413 (16).

High-resolution mass spectroscopy HR-ESI-MS: m / z: 523.2671 [M+Na] ⁺ (calc. C ₂₉ H ₄₀ O ₇ Na, 523.2672).

Regarding the physicochemical properties of the compound Anomanolide A (19):

Melting point (mp): 170~172 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 214 nm; infrared absorption spectrum (IR): ν _max 3432, 2952, 1720, 1675, 1380, 1128, 754 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, CDCl ₃ ): δ 6.19 (1H, d, J = 10.0 Hz, H-2 ), 6.93 (1H, dd, J = 10.0, 6.0 Hz, H-3 ), 3.76 (1H, d, J = 6.0 Hz, H-4 ), 3.24 (1H, s(br), H-6 ), 1.35 (1H, dd, J = 15.2, 3.2 Hz, H-7a ), 2.16 (1H, ddd, J = 15.2,4.0,3.2Hz, H-7b), 1.55 (1H, td, J = 10.8,4.0Hz, H-8), 1.01 (1H, td, J = 10.8,3.2Hz , H-9 ), 1.37 (1H, m, H-11a ), 1.92 (1H, m, H-11b ), 1.42 (2H, m, H-12a, b ), 1.43 (1H, m, H-14 ), 1.23 (1H, m, H-15a ), 1.73 (1H, m, H-15b ), 1.70 (1H, m, H-16a ), 2.04 (1H, d, J = 8.4 Hz, H-16b ) , 0.75 (3H, s, CH ₃ -18 ), 1.40 (3H, s, CH ₃ -19 ), 2.43 (1H, ddd, J = 9.2, 7.6, 1.0 Hz, H-20 ), 1.31 (1H, dd , J = 12.8, 7.6 Hz, H-21a ), 2.51 (1H, ddd, J = 12.8, 9.2, 2.0 Hz, H-21b ), 4.66 (1H, d, J = 2.4 Hz, H-22 ), 2.07 (1H,d, J = 13.2Hz, H-23a ), 1.78 (1H, dd, J = 13.2, 2.4Hz, H-23b ), 1.45 (3H, s, CH ₃ -27 ), 1.18 (3H, s , CH ₃ -28 ).

Mass spectrum FAB-MS m / z (rel.int.): 487 [M+H] ⁺ .

High-resolution mass spectroscopy HR-ESI-MS: m / z: 509.2517 [M+Na] ⁺ (calc. C ₂₈ H ₃₈ O ₇ Na, 509.2510).

Regarding the physicochemical properties of the compound Anomanolide B (20):

Melting point (mp): 168-170 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 214, 230 nm; infrared absorption spectrum (IR): ν _max 3448, 2960, 1726, 1675, 1369, 1130 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.98 (1H, dd, J = 10.0, 2.0 Hz, H-2 ), 6.45 (1H, dd, J = 10.0, 2.0 Hz, H -3 ), 5.03 (1H, s(br), H-4 ), 4.42 (1H, dd, J = 12.8, 4.4 Hz, H-6 ), 1.72 (1H, m, H-7a ), 2.32 (1H , 2.32dt, J = 9.6, 4.4Hz, H-7b ), 1.65 (1H, m, H-8 ), 1.28 (1H, dd, J = 9.6, 3.2Hz, H-9 ), 0.96(1H,d , J = 8.0 Hz, H-11a ), 1.32 (1H, m, H-11b ), 1.35 (1H, m, H-12a ), 1.39 (1H, m, H-12b ), 1.59 (1H, m, H-14 ), 1.27 (1H, m, H-15a ), 1.73 (1H, m, H-15b ), 1.66 (1H, m, H-16a ), 2.08 (1H, m, H-16b ), 0.72 (3H, s, CH ₃ -18 ), 1.24 (3H, s, CH ₃ -19 ), 2.40 (1H, q, J = 7.6 Hz, H-20 ), 1.25 (1H, m, H-21a ), 2.44 (1H, dd, J = 12.8, 2.4 Hz, H-21b ), 4.64 (1H, d, J = 2.4 Hz, H-22 ), 2.05 (1H, d, J = 14.8 Hz, H-23a ), 1.74 (1H, dd, J = 14.8,2.4Hz, H-23b), 1.43 (3H, s, CH 3 -27), 1.15 (3H, s, CH 3 -28).

Mass Spectrum ESI-MS m / z ( s. s.): 505 [M+H] ⁺ .

About the physicochemical properties of the compound Anomanolide C (21):

Melting point (mp): 280-282 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 214 nm; infrared absorption spectrum (IR): ν _max 3430, 2927, 1702, 1677, 1369, 1130 cm ^-1 Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.38 (1H, d, J = 10.0 Hz, H-2 ), 7.18 (1H, dd, J = 10.0, 6.4 Hz, H- 3 ), 3.98 (1H, dd, J = 6.4, 2.4 Hz, H-4 ), 3.16 (1H, s(br), H-6 ), 1.20 (1H, dd, J = 14.0, 11.2 Hz, H- 7a ), 2.04 (1H, m, H-7b ), 1.51 (1H, m, H-8 ), 1.02 (1H, ddd, J = 11.2, 4.0, 2.8 Hz, H-9 ), 1.51 (1H, m , H-11a), 2.04 ( 1H, m, H-11b), 1.58 (1H, m, H-12a), 2.01 (1H, m, H-12b), 1.98 (1H, m, H-14), 1.67 (1H, m, H-15a ), 1.75 (1H, dd, J = 13.6, 7.2 Hz, H-15b ), 4.51 (1H, dd, J = 7.2, 3.2 Hz, H-16 ), 0.63 (3H) , s, CH ₃ -18 ), 1.80 (3H, s, CH ₃ -19 ), 2.75 (1H, dd, J = 8.4, 8.0 Hz, H-20 ), 1.98 (1H, m, H-21a ), 2.99 (1H, dd, J = 9.6, 8.0 Hz, H-21b ), 5.47 (1H, d(br), J = 2.0 Hz, H-22 ), 2.15 (1H, d, J = 12.8 Hz, H- 23a), 2.04 (1H, dd , J = 12.8,2.8Hz, H-23b), 1.67 (3H, s, CH 3 -27), 1.34 (3H, s, CH 3 -28).

Mass spectrum EI-MS m / z (rel. int): 502 [M] ⁺ .

High resolution mass spectrum HR-ESI-MS: m / z 525.2466 [M + Na] + ( calcd for _{C 28 H 38 O 8 Na,} 525.2459).

Regarding the physicochemical properties of the compound Anomanolide D (22):

Melting point (mp): 196~198 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 214 nm; infrared absorption spectrum (IR): ν _max 3448, 2948, 1718, 1675, 1378, 1126, 1049 cm ^-1 ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.16 (1H, dd, J = 10.0, 2.0 Hz, H-2 ), 6.77 (1H, dd, J = 10.0, 2.0 Hz, H-3 ), 5.19 (1H, s(br), H-4 ), 4.64 (1H, dd, J = 12.8, 4.4 Hz, H-6 ), 1.82 (1H, t, J = 12.0 Hz, H-7a ), 2.21 (1H, dt, J = 12.8, 4.0 Hz, H-7b ), 1.59 (1H, m, H-8 ), 1.59 (1H, m, H-9 ), 1.10 (1H, m , H-11a ), 1.37 (1H, m, H-11b ), 1.46 (1H, dt, J = 9.6, 2.8 Hz, H-12a ), 1.94 (1H, m, H-12b ), 2.12 (1H, m, H-14 ), 1.72 (2H, d, J = 11.2 Hz, H-15a, b ), 4.50 (1H, d(br), J = 3.2 Hz, H-16 ), 0.61 (3H, s, CH ₃ -18 ), 1.53 (3H, s, CH ₃ -19 ), 2.71 (1H, t, J = 8.4 Hz, H-20 ), 1.93 (1H, m, H-21a ), 2.95 (1H, dd , J =11.2, 9.6 Hz, H-21b ), 5.45 (1H, d, J = 2.0 Hz, H-22 ), 2.14 (1H, d, J = 12.8 Hz, H-23a ), 2.02 (1H, dd , J = 12.8, 3.2 Hz, H-23b ), 1.66 (3H, s, CH ₃ -27 ), 1.34 (3H, s, CH ₃ -28 ).

Mass spectrum FAB-MS m / z ( rel.int.): 539 [M+H] ⁺ .

High resolution mass spectrum HR-ESI-MS: m / z 561.2229 [M + Na] + ( calcd for _{C 29 H 39 ClO 8 Na,} 561.2226).

Regarding the physicochemical properties of the compound Anomanolide E (23):

Melting point (mp): 182-184 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 214 nm; infrared absorption spectrum (IR): ν _max 3455, 2933, 1720, 1675, 1400, 1132 cm ^-1 Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.12 (1H, dd, J =10.0, 2.4 Hz, H-2 ), 6.64 (1H, ddd, J =10.0, 3.6, 2.4 Hz, H-3 ), 2.39 (1H, d, J = 19.2 Hz, H-4eq ), 3.73 (1H, ddd, J = 19.6, 2.4, 2.4 Hz, H-4ax ), 4.10 (1H, s (br) ), H-6 ), 1.85 (1H, dt, J = 12.0, 2.8 Hz, H-7a ), 2.23 (1H, td, J = 12.0, 2.4 Hz, H-7b ), 2.14 (1H, m, H -8 ), 2.56 (1H, td, J = 11.2, 3.2 Hz, H-9 ), 1.58 (1H, qd, J = 12.0, 3.2 Hz, H-11a ), 2.83 (1H, dt, J = 12.0, 3.2 Hz, H-11b ), 1.71 (1H, m, H-12a ), 2.41 (1H, m, H-12b ), 2.41 (1H, m, H-14 ), 1.79 (2H, m, H-15a , b ), 4.49 (1H, d, J = 6.4 Hz, H-16 ), 0.76 (3H, s, CH ₃ -18 ), 1.67 (3H, s, CH ₃ -19 ), 2.79 (1H, dd, J = 8.4, 8.0 Hz, H-20 ), 2.04 (1H, dd, J = 12.8, 7.6 Hz, H-21a ), 3.02 (1H, ddd, J = 12.8, 9.6, 1.2 Hz, H-21b ), 5.47 (1H, d, J = 2.0 Hz, H-22 ), 2.15 (1H, d, J = 12.0 Hz, H-23a ), 2.07 (1H, dd, J = 12.0, 2.8 Hz, H-23b ), 1.67 (3H, s, CH ₃ -27 ), 1.36 (3H, s, CH ₃ -28 ).

Mass spectrum FAB-MS m / z (rel. int): 505 [M+H] ⁺ .

High-resolution mass spectroscopy HR-ESI-MS: m / z 527.2618 [M+Na] ⁺ (calc. C ₂₈ H ₄₀ O ₈ Na, 527.2615).

About the physicochemical properties of the compound Anomanolide F (24):

Melting point (mp): 190-192 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 215 nm; infrared absorption spectrum (IR): ν _max 3416, 2929, 2863, 1712, 1668, 1376, 1082 , 753 cm ^-1 ; hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.12 (1H, dd, J = 10.4, 2.4 Hz, H-2 ), 6.73 (1H, dd, J = 10.4 , 2.4 Hz, H-3 ), 5.35 (1H, br s, H-4 ), 1.22 (1H, m, H-6a ), 2.22 (1H, m, H-6b ), 1.24 (1H, m, H -7a ), 1.78 (1H, m, H-7b ), 1.65 (1H, m, H-8 ), 1.56 (1H, m, H-9 ), 1.38 (2H, m, H-11ab ), 1.54 ( 1H, m, H-12a ), 1.99 (1H, m, H-12b ), 2.16 (1H, m, H-14 ), 2.09 (2H, m, H-15ab ), 4.42 (1H, s(br) , H-16 ), 0.71 (3H, s, CH ₃ -18 ), 1.61 (3H, s, CH ₃ -19 ), 2.55 (1H, t, J = 8.8, 8.4 Hz, H-20 ), 1.96 ( 1H, m, H-21a ), 2.71 (1H, d, J = 12.0 Hz, H-21b ), 5.00 (1H, m, H-22 ), 2.02 (1H, m, H-23a ), 2.05 (1H , m, H-23a ), 1.81 (3H, s, CH ₃ -27 ), 1.40 (3H, s, CH ₃ -28 ).

Mass Spectrum ESI-MS m / z ( s. s.): 527 [M+Na] ⁺ , 527 (100), 413 (61).

High resolution mass spectrum HR-ESI-MS: m / z 527.2620 [M + Na] + ( calcd for _{C 28 H 40 O 8 Na,} 527.2621).

About the physicochemical properties of the compound Tubonolide A (25):

Melting point (mp): 200~202 ° C; ultraviolet absorption spectrum (UV, in methanol solution): λ _max 214 nm; infrared absorption spectrum (IR): ν _max 3427, 2938, 1730, 1676, 1371, 985, 752 cm ^{- 1} ; Hydrogen nuclear magnetic resonance spectrum ¹ H NMR (400 MHz, C ₅ D ₅ N ): δ 6.25 (1H, dd, J = 8.0, 1.2 Hz, H-2 ), 6.83 (1H, dd, J = 8.0, 2.0 Hz , H-3 ), 5.23 (1H, m(br), H-4 ), 4.69 (1H, dd, J = 10.0, 4.0 Hz, H-6 ), 1.88 (1H, dd, J = 10.0, 10.0 Hz) , H-7a ), 2.26 (1H, ddd, J = 10.0, 4.0, 3.2 Hz, H-7b ), 1.59 (1H, m, H-8 ), 1.56 (1H, td, J = 8.8, 3.2 Hz, H-9 ), 1.09 (1H, m, H-11a ), 1.43 (1H, m, H-11b ), 1.41 (1H, m, H-12a ), 1.83 (1H, m, H-12b ), 2.13 (1H, m, H-14 ), 1.72 (1H, m, H-15a ), 1.79 (1H, m, H-15b ), 4.49 (1H, t, J = 4.4 Hz, H-16 ), 0.75 ( 3H, s, CH ₃ -18 ), 1.62 (3H, s, CH ₃ -19 ), 2.62 (1H, m, H-20 ), 2.67 (1H, dd, J = 10.4, 5.6 Hz, H-21ax ) , 1.64 (1H, d, J = 10.4 Hz, H-21eq ), 5.09 (1H, s(br), H-22 ), 2.91 (1H, d, J = 12.4Hz, H-23ax ), 2.15 (1H , m, H-23b ), 1.50 (3H, s, CH ₃ -27 ), 1.36 (3H, s, CH ₃ -28 ), 7.43 (1H, d, J = 5.6 Hz, 4-OH ), 5.99 ( 1H, s, 5-OH ), 7.28 (1H, d, J = 3.6 Hz, 16-OH ), 5.41 (1H, s, 17-OH ), 6.11 (1H, s, 24-OH ).

Mass Spectrum ESI-MS m / z ( s. s.): 539 [M+H] ⁺ .

High resolution mass spectrum HR-ESI-MS: m / z 561.2232 [M + Na] + ( calcd for _{C 28 H 39 ClO 8 Na,} 561.2226).

Biological activity test

The present invention further performs the biological activity test of the twenty-five new withanolides obtained. The biological activity assay of the present invention uses five human cancer cell lines, including two human breast cancer cell lines (MCF-7 and MDA-MB-231), two human liver cancer cell lines (Hep G2 and Hep 3B), and a Human lung cancer cell line (A549). In addition, a human lung cell (MRC-5) was used as a normal cell control group. The human cancer cell lines were all from the American Type Culture Collection, and each of the cells was cultured in 10% (v/v) fetal calf serum (100%/ml of penicillin) (penicillin). ) in a RPMI-1640 suspension with 100 g/ml streptomycin and maintained at 37 ° C in an environment containing 5% carbon dioxide and 95% air.

In the present invention, the cytotoxicity test is carried out by a well-known method of MTT (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide). Five cancer cell lines as well as normal cell lines (MRC-5) were planted in 96-well tissue culture dishes at a density of 5,000-10,000 cells/well, respectively. On the next day, cells were treated with the test compound (AD) and Doxorubicin (Compound E), and culture was continued for 72 hours, after which the cells were tested by MTT method and dissolved in DMSO. The absorbance of the test results was analyzed at 550 nm using a microplate reader to examine the toxicity of each compound to each cell line. The results of the biological test are shown in Table 1, which is the toxic effect of each compound on the cancer cell line, wherein the IC ₅₀ value represents the concentration of the compound which inhibits 50% of cell growth, and the Doxorubicin (Compound E) having the activity of killing cancer cells As a positive control. As shown in Table 1, in the results of poisoning the cancer cell lines of each compound, it was found that the compounds 1 , 8, 9, 10, and 14 had good toxicity to the above five cancer cell lines, and the IC _{50 thereof was} less than 2.0 μg/ml. .

In summary, the "composition for killing cancer cells and preparation method thereof" in the present case can not only obtain new compounds withanolides from the natural plant Dragon Ball ( Tubocapsicum anomalum ), but more particularly The novel asanolides compounds have the biological activity of killing cancer cells. However, the embodiments described in the above description are merely illustrative of the principles of the invention and its effects, and are not intended to limit the invention. Therefore, those skilled in the art can make modifications and changes to the above embodiments without departing from the spirit of the invention. The scope of the invention should be as set forth in the appended claims.

Claims (6)

A composition for killing cancer cells comprising a compound of the form of the formula (withanolides) of the formula: Wherein the cancer cell line is selected from the group consisting of human liver cancer cells, human breast cancer cells, human lung cancer cells, and combinations thereof. The composition of claim 1, wherein the withanolides compound is obtained by extracting from a Solanaceae plant. The composition of claim 2, wherein the Solanaceae plant is Tubocapsicum anomalum . A use of a compound comprising the following structure for preparing an anticancer drug for use in poisoning at least one of human liver cancer cells and human breast cancer cells The use of claim 4, wherein the compound is extracted from a Solanaceae plant. The use of the fifth aspect of the patent application, wherein the Solanaceae plant is Tubocapsicum anomalum .